GTG Banding Research Articles

T (1;12) (p31; q13) cytogenetic abnormality in a patient of idiopathic hypereosinophilic syndrome: A case report

Hypereosinophic syndrome (HES) is a heterogenous group of disorders comprising various etiology. Idiopathic HES is diagnosis of exclusion after morphological, radiological and molecular investigations. Cytogenetic analysis remains an important diagnostic test available in HES. The patient reported here presented with hyper eosinophilia with cardiac, gastrointestinal and pulmonary organ involvement. Chromosomal abnormality of t (1;12) (p31; q13) was found in conventional karyotyping (unstimulated culture, GTG banding) on bone marrow sample. Patient improved after short course of corticosteroid and imatinib mesylate a tyrosine kinase inhibitor (TKI).

Donor Screening: Level of Sensitivity in Karyotype with GTG banding vs PGT-A with NGS

Donor Screening: Level of Sensitivity in Karyotype with GTG banding vs PGT-A with NGS

New Karyotype Information for Ctenomys (Rodentia: Ctenomyidae) from Midwest and Northern Brazil

Introduction: Its wide karyotypic variation characterizes the genus Ctenomys, and in Brazil, the genus is distributed in the country’s southern, Midwest, and northern regions. Recently, populations of Ctenomys have been found in the Midwest and northern Brazil, with two new lineages named C. sp. “xingu” and C. sp. “central.” Methods: This work combines classical cytogenetic and molecular analyses to provide new chromosomal information on the boliviensis group distributed in northern and Midwestern Brazil. This includes the validation of the karyotype of C. bicolor and C. nattereri and the description of the karyotype of C. sp. “xingu” and C. sp. “central.” Results: We found three different karyotypes: 2n = 40 for C. bicolor; 2n = 36 for C. nattereri, and specimens from a locality belonging to C. sp. “central”; 2n = 34 for the lineage C. sp. “xingu” and specimens from a locality belonging to C. sp. “central.” Furthermore, GTG banding revealed homologous chromosomes between species/lineages and allowed the identification of the rearrangements that occurred, which proved the occurrence of fissions. Conclusion: Considering our results on the variation of 2n in the boliviensis group, we found two possibilities: the first, deduced by parsimony, is that 2n = 36 appeared initially, and two fissions produced gave rise to 2n = 40, and an independent fusion gave rise to 2n = 34 from 2n = 36; moreover, the second explanation is that all karyotypes arose independently.

Cytogenetic abnormalities associated with reproductive failure in Pakistani population: Experience of a tertiary care hospital.

Constitutional chromosomal abnormalities play a significant role in causing reproductive anomalies in individuals of reproductive age. With the rapid advancement of genome engineering techniques, it has now become possible to cure different genetic disorders. However, very limited data is available regarding the prevalence of such aberrations in the Pakistani population. Considering this factor, this retrospective analysis was undertaken to elucidate the type and prevalence rate of such abnormalities in our population. A total of 241 individuals, who were referred to the Liaquat National Hospital, from January 2017 to December 2021, with a history of infertility or miscarriages, were evaluated using the standard GTG banding technique. The results revealed a notably high percentage 44(18.2%) of chromosomal abnormalities in our population. Surprisingly, the frequency of these anomalies was observed to be higher in males than in females. However, further research is needed using a larger sample size to confirm the findings of this investigation.

Biotechnological aspects of the working-out and manufacturing of living bone equivalent

Objective. To handle biotechnological aspects in manufacturing processes of three-dimensional living bone equivalent for restoration of critical sized bone defects for innovative treatment of combat-related casualties. Methods. To fabricate living bone equivalent we used devitalized xenogeneic bone scaffolds (DBM chips) and autologous fibrin hydrogel seeded with autologous cultured bone marrow-derived multipotent mesenchymal stem/stromal cells (BM-MSCs). Quality/identity control of cell cultures was assured by donor and cell culture infection screening (IFA, PCR), flow cytometry (cell phenotype), karyotyping (GTG banding), functional assays (CFU assay, multilineage differentiation assay). Results. The BM-MSC cultures had a normal karyotype and appropriate phenotype, multilinear differentiation potential and functional properties, appropriate CFU frequency and hadn’t any signs of cell senescence. The FDA/PI combined staining showed the demineralized bone chips’ regular seeding with viable cells. Conclusions. An actual regenerative medicine approach to organ-saving transplantation of the three-dimensional living bone equivalent for combat-related casualties requires further preclinical and clinical approbation for thorough studies on the bone integrity restoration, forming new bone tissue in a site of bone defect, and duration of rehabilitation period compared to the gold standard of the conventional bone defect cure.

Chromosomal abnormalities in children with acute lymphoblastic leukemia

Acute lymphoblastic leukemia is the most conventional cancer in children and teenagers. In adults, this pathology is much less common. The paper presents the results of a retrospective analysis of chromosomal aberrations in clinically identified 1269 feeble children with acute lymphoblastic leukemia for the period from January 2013 to January 2023. The average age of the patients was 10.2±1.69 years. The material for the study was bone marrow and blood cells obtained from a puncture biopsy of the sternum and peripheral vein. Cytogenetic studies were carried out using the generally accepted standard method using GTG banding. The results of the analysis showed the absence of genomic and chromosomal mutations in 215 (17%) children; translocation t(1;19)(q23;p13) was detected in 75 (6%) children; in 221 (17.5%) children - t(4;11)(q21;q23); in 234 (18.5%) children - t(9;22)(q34;q11); 88 (7%) - t(12;21)(p33;q22) and 139 (11%) patients had translocations between other chromosomes. Hyperdiploid translocations were detected in 139 (11%) children. No sick children with hypodiploid cells were found. No metaphase cells were obtained from 152 patients (12%). In sick children, a relatively low frequency of optimistic predicting chromosomal abnormalities, namely t(12;21)(p13;q22) and hyperdiploid, has been established. The pervasiveness of chromosomal aberrations with a negative treatment prediction, such as t(9;22)(q34;q11.2), is consistent with the accessible international scientific literature data. The results obtained indicate the importance of cytogenetic studies in the diagnosis and predictions of children with ALL.

A retrospective analysis of spontaneous chromosomal aberrations in human lymphocyte cultures of individuals from Bosnia and Herzegovina

Spontaneous chromosomal aberrations are structural or numerical changes of chromosomes that occur naturally, without exposure to external genotoxic factors. They are not inherited, occur randomly in the karyotype, and do not have direct clinical significance. However, they can affect genomic instability and disease predisposition. They can result from DNA replication or repair processes errors, and typically are observed in cells that are actively dividing. Spontaneous chromosomal aberrations may arise due to the natural chromosomal instability and can be elevated in individuals exposed to mutagens. We analyzed frequencies of spontaneous chromosomal aberrations in 137 individuals subjected to karyotype analysis at the Laboratory for Cytogenetics and Genotoxicology, University of Sarajevo – Institute for Genetic Engineering and Biotechnology, during 2008-2023. Whole blood samples were cultivated for 72 hours with the thymidine added in the 48th hour. Metaphases were arrested by colcemid 60 minutes before harvesting. GTG banding was performed and slides were analyzed under 1000x magnification in accordance with An International System for Human Cytogenetic Nomenclature and E.C.A. Cytogenetic Guidelines and Quality Assurance. Constitutionally aberrant karyotypes were found in 2.92% of analysed individuals as well as altered karyotypes considered as normal chromosomal variants. In the total of 3092 analyzed metaphases, 20 spontaneous chromosomal aberrations were found in 13 individuals. This study contributes to the limited knowledge of the cytogenetic status of the Bosnian and Herzegovinian population. Further monitoring of spontaneous chromosomal aberrations incidences is recommended.

A genotype-phenotype correlation in split-hand/foot malformation type 1: further refinement of the phenotypic subregions within the 7q21.3 locus.

Background: Split-hand/foot malformation type 1 (SHFM1) refers to the group of rare congenital limb disorders defined by the absence or hypoplasia of the central rays of the autopods with or without accompanying anomalies, such as hearing loss, craniofacial malformation, and ectodermal dysplasia. Consequently, the condition is characterized by clinical variability that hinders diagnostic and counseling procedures. SHFM1 is caused by pathogenic variants affecting the DLX5/6 genes and/or their tissue-specific enhancers at the 7q21.3 locus. Herein, we report on seven patients from five unrelated Polish families affected by variable symptoms of the SHFM1 spectrum, all harboring 7q21.3 or 7q21.2-q21.3 rearrangements, and provide a genotype-phenotype correlation in the studied cohort. Methods: We applied GTG banding, array-based comparative genomic hybridization (aCGH), and whole-genome sequencing (WGS) in order to identify the causative aberrations in all affected patients. Results: The identified pathogenic structural variants included deletions and/or translocations involving the 7q21.3 locus, i.e., t(7;10)(q21.3;q22.2) and t(7;12)(q21.3;q21.2) in all affected individuals. Interestingly, a sporadic carrier of the latter aberration presented the SHFM1 phenotype with additional features overlapping with Baker-Gordon syndrome (BAGOS), which resulted from the translocation breakpoint at chromosome 12 within the SYT1 gene. Conclusion: Clinical variability of the studied cohort reflects the composition of the DLX5/6 regulatory elements that were dislocated from their target genes by chromosomal rearrangements. The correlation of our data with the previously published observations enabled us to update the phenotypic subregions and regulatory units within the SHFM1 locus. In addition, we present the first case of SHFM1 and BAGOS-like phenotype that resulted from translocation breakpoints at chromosomes 7 and 12, both of which were pathogenic, and consequently, we show the first evidence that BAGOS can also result from the regulatory loss-of-function SYT1 mutations. In this paper, we emphasize the utility of sequence-based approaches in molecular diagnostics of disorders caused by regulatory structural variants.

Correlation between conventional karyotype, interphase FISH and immunophenotype in risk stratification of newly diagnosed plasma cell myeloma: An observational study

BackgroundPlasma cell myeloma (PCM) is a common adult hematological neoplasm of terminally differentiated B-cells resulting in accumulation of monoclonal plasma cells. PCM is heterogeneous disease with survival time varies from months to years, determined by age, stage, cytogenetics abnormalities, and treatment response. There is conflicting evidence in role of immunophenotype as a prognostic indicator. This study identified frequency of cytogenetics abnormalities and established correlation between cytogenetics and immunophenotyping in risk stratification in newly diagnosed PCM. MethodsPilot study was undertaken on 30 newly diagnosed PCM over 4 years. Apart from extensive myeloma laboratory and imaging profile, multicolor flow cytometry using panel of CD138, CD38, CD45, CD56, CD19, CD20, CD28, CD200, CD117, κ, and λ light-chains. Karyotyping was done using GTG banding and interphase fluorescence in situ hybridization analysis on magnetically sorted plasma cells for t(4;14), t(11;14), del(13)q, and del(17)p probes. ResultsOf 30 myeloma patients, 16(53.3%) were men with mean age 57.23 ± 11.90 years. The del(13)q was commonest cytogenetics abnormality. CD56-positive cases were strongly associated with presence of del(17)p and stage III of disease (p < 0.05). Negative expression of CD117 was seen in advanced stage of disease and high-risk genetic factor, t(4;14) (p = 0.049). Absent CD28 expression was associated with higher albumin levels, lower levels of β2MG, and lack of high-risk genetic abnormalities. Advanced clinical stages were associated with expression of CD56, CD28, and absent expression of CD45 and CD117. ConclusionImmunophenotypic antigens positive and negative expressions are associated with high-risk genetic abnormalities and advanced clinical stages of PCM. Institution ethic committee reference numberIEC/2017/230, AFMC, Pune.

Cytogenetic study for Iraqi patients with epilepsy

The current study was conducted with the aim of identifying the chromosomal changes that associated with epilepsy Iraqi patients. Chromosomal analysis was carried on for 37 of a number of epilepsy patients exclusively for cases of idiopathic epilepsy (unknown cause)after excluding the symptomatic epilepsy (known causes) range age from 2-51 years. Only five cases showed chromosomal changes associated with epilepsy cases, their ages ranged from 2-30 years, while the rest of the patients had normal karyotype. The chromosomes were prepared using the GTG band with average of 25-30 cells was examined for each person. The chromosomal changes that associated with epilepsy cases, ranged from the presence of a number of derivative chromosomes, inversion, additions, deletions, chromatid fractures, and the premature centromere division (PCD).Finally the recurrence of aberrations in some chromosomes (more than one defect associated with the same chromosome). Which may indicate that these chromosomes carry more than one fragile site sin same chromosome. During questionnaire there were other symptoms associated with epilepsy which has been mentioned to them in the study.

Trisomy 5: A Rare Isolated Finding in Pediatric B-lymphoblastic Leukemia

Background and Objectives: In acute leukemia, all diagnostic criteria and treatment protocols are based on cytogenetic and molecular genetic findings. Despite recent advances in molecular biology, cytogenetic studies still play a pivotal role in the sub-classification of B-lymphoblastic leukemia (B-ALL). B-ALL is characterized by clonal cytogenetic abnormalities with numerical chromosomal aberrations being more common. An extra copy of chromosome 5 is common to see in cases with hyper diploidy. However, a gain of chromosome 5 as a sole anomaly is exceptionally rare in B-ALL. To date, trisomy 5 as a sole abnormality is reported in few cases only. We aimed to report the clinicopathologic profile of this rare finding to increase knowledge and highlight the disease course of these patients.&#x0D; Methods: We report a case of a 14-year boy presented with fever, lethargy and episodes of nasal bleeding for two weeks. He was admitted to the pediatric oncology unit at Indus hospital and health network, Karachi. Flowcytometry performed on peripheral blood using 8-color flowcytometry. Conventional karyotyping was performed by GTG banding. FISH panel was comprised of dual color dual fusion probes for BCR::ABL1 and ETV6::RUNX1 whereas break apart probe for KMT2A (Metasystem, Germany). Digital image analysis for karyotyping and FISH was done on Leica Biosystems, Cytovision MB8.&#x0D; Results: Flowcytometry results were consistent with B-ALL. Cytogenetic analysis on his bone marrow aspirate revealed trisomy 5 as a sole abnormality with no evidence of any other clonal cytogenetic abnormality. FISH studies for BCR::ABL1, ETV6::RUNX1 and KMT2A showed no evidence of gene rearrangements.&#x0D; Conclusion: Trisomy 5 is a very rare cytogenetic aberration. Only few cases reported in children. Inferior outcome is reported in both children and adults. The increasing number of reported cases raises the possibility of a distinct cytogenetic entity. Its prognostic and therapeutic implications are yet to be explored.

Characterization of the chromosomes of sotol (Dasylirion cedrosanum Trel.) using cytogenetic banding techniques

Sotol (Dasylirion cedrosanum Trel.) is a perennial species with numerous grayish-to-green leaves that grow symmetrically from the base of the stem outward. Its inflorescences can measure up to 3 m in height and contain membranous bracts that enclose seeds. It is a species that has been scarcely studied at the cytogenetic level, with only one report available in the literature. In Mexico, it has economic importance because it is used to prepare the alcoholic beverage sotol. In the present work, the chromosomes of Dasylirion cedrosanum Trel. were obtained and analyzed using different cytogenetic banding techniques and morphometric analysis to construct the first karyotype for this species. Chromosomes were obtained by germinating plant seeds collected in the locality of Las Adjuntas, Santiago, Nuevo León, Mexico. Treatment with colchicine as an antimitotic was performed, followed by enzymatic treatment with pectinase and cellulase, to eliminate the cell walls. Chromosome slides were stained with Giemsa, GTG banding technique, CBW banding, and the 4’,6-diamidino-2-phenylindole fluorescence dye, and observed under a microscope. A chromosomal number 2n of 38 chromosomes, as previously reported, was confirmed. Using the different banding techniques, we observed that all chromosomes exhibited a submetacentric morphology with a fundamental number of 76, and it was possible to visualize the pattern of GTG and CBW bands; these findings are reported for the first time for this species. Morphometric analysis established that the average length of the chromosomes was between 5.09 and 9.84 mm.

Cytogenetic Analysis of Sporadic First-Trimester Miscarriage Specimens Using Karyotyping and QF-PCR: A Retrospective Romanian Cohort Study.

It is well known that first-trimester miscarriages are associated with chromosome abnormalities, with numerical chromosome abnormalities being the ones most commonly detected. Conventional karyotyping is still considered the gold standard in the analysis of products of conception, despite the extended use of molecular genetic techniques. However, conventional karyotyping is a laborious and time-consuming method, with a limited resolution of 5-10 Mb and hampered by maternal cell contamination and culture failure. The aim of our study was to assess the type and frequency of chromosomal abnormalities detected by conventional karyotyping in specimens of sporadic first-trimester miscarriages in a Romanian cohort, using QF-PCR to exclude maternal cell contamination. Long-term cultures were established and standard protocols were applied for cell harvesting, slide preparation, and GTG banding. All samples with 46,XX karyotype were tested for maternal cell contamination by QF-PCR, comparing multiple microsatellite markers in maternal blood with cell culture and tissue samples. Out of the initial 311 specimens collected from patients with sporadic first-trimester miscarriages, a total of 230 samples were successfully analyzed after the exclusion of 81 specimens based on unsuitable sampling, culture failure, or QF-PCR-proven maternal cell contamination. Chromosome abnormalities were detected in 135 cases (58.7%), with the most common type being single autosomal trisomy (71/135-52.6%), followed by monosomy (monosomy X being the only one detected, 24/135-17.8%), and polyploidy (23/135-17.0%). The subgroup analysis based on maternal age showed a statistically significant higher rate of single trisomy for women aged 35 years or older (40.3%) compared to the young maternal age group (26.1%) (p = 0.029). In conclusion, the combination of conventional karyotyping and QF-PCR can lead to an increased chromosome abnormality detection rate in first-trimester miscarriages. Our study provides reliable information for the genetic counseling of patients with first-trimester miscarriages, and further large-scale studies using different genetic techniques are required.

Investigation of the Frequency and Type of Chromosomal Abnormalities in Women Patients with Amenorrhea.

Amenorrhea is defined as the absence of menstruation at the reproductive age of women. Amenorrhea caused by various etiological factors including genetic factors, intrauterine malformations, endocrine dysfunction, and environmental factors. Genetic factors particularly chromosomal abnormalities are the main cause of Amenorrhea. This study was performed to estimate the frequency and types of chromosomal abnormalities in patients with amenorrhea in the northeast of Iran. A total of 381 women with the history of amenorrhea participated in this study. Peripheral blood lymphocyte cultures were performed according to the standard GTG banding method. 296 (77%) of a total of all cases had a normal karyotype (46, XX) while 85 patients (23%) had abnormal karyotype. The numerical and structural abnormalities of X chromosome were observed in 52 (61%), the abnormalities of Y chromosome were observed in 23 (27.2%) and rearrangements between autosomal and/or sex chromosomes were observed in 10 (11.8%). The present study revealed that cytogenetic study is essential for early diagnosis and treatments of Amenorrhea.

Candidate genes for infertility: an in-silico study based on cytogenetic analysis

BackgroundThe cause of infertility remains unclear in a significant proportion of reproductive-age couples who fail to conceive naturally. Chromosomal aberrations have been identified as one of the main genetic causes of male and female infertility. Structural chromosomal aberrations may disrupt the functioning of various genes, some of which may be important for fertility. The present study aims to identify candidate genes and putative functional interaction networks involved in male and female infertility using cytogenetic data from cultured peripheral blood lymphocytes of infertile patients.MethodsKaryotypic analyses was done in 201 infertile patients (100 males and 101 females) and 201 age and gender matched healthy controls (100 males and 101 females) after 72 h peripheral lymphocyte culturing and GTG banding, followed by bioinformatic analysis using Cytoscape v3.8.2 and Metascape.ResultsSeveral chromosomal regions with a significantly higher frequency of structural aberrations were identified in the infertile males (5q2, 10q2, and 17q2) and females (6q2, 16q2, and Xq2). Segregation of the patients based on type of infertility (primary v/s secondary infertility) led to the identification of chromosomal regions with a significantly higher frequency of structural aberrations exclusively within the infertile males (5q2, 17q2) and females (16q2) with primary infertility. Cytoscape identified two networks specific to these regions: a male specific network with 99 genes and a female specific network with 109 genes. The top enriched GO terms within the male and female infertility networks were “skeletal system morphogenesis” and “mRNA transport” respectively. PSME3, PSMD3, and CDC27 were the top 3 hub genes identified within the male infertility network. Similarly, UPF3B, IRF8, and PSMB1 were the top 3 hub genes identified with the female infertility network. Among the hub genes identified in the male- and female-specific networks, PSMB1, PSMD3, and PSME3 are functional components of the proteasome complex. These hub genes have a limited number of reports related to their respective roles in maintenance of fertility in mice model and humans and require validation in further studies.ConclusionThe candidate genes predicted in the present study can serve as targets for future research on infertility.

PB1891: VARIANT PHILADELPHIA CHROMOSOME IN CML

Background: Chronic myeloid leukemia is a clonal malignant disorder of a pluripotent hematopoietic stem cell characterized by the Philadelphia chromosome and the reciprocal translocation t(9:22)(q34;q11). Almost 5 to 10 percent of chronic myeloid leukemia cases demonstrate variant Philadelphia translocations. Aims: To contribute to the description of patients’ clinical characteristics and prognosis with variant Ph chromosome-positive leukemia, we report a Ph chromosome-positive CML in the chronic phase characterized by an unusual variant Ph chromosome. Methods: An elderly male Libyan patient was suspected to have chronic myeloid leukemia. Cytogenetic and molecular exploration were conducted using bone marrow and blood samples. Cytogenetic analysis was performed on lymphocytes and bone marrow cells after short-term culture (48 hours). The cells were treated with colchicine and hypotonic KCl solution. The pellet was fixed and washed in methanol-acetic acid (3:1). The cells were resuspended in fixative and dropped onto slides. Chromosome-banding analysis was performed using RHG and GTG banding techniques. Three karyotypes and 25 metaphases were analyzed according to the International System for Human Cytogenetic Nomenclature. Multiplex reverse transcription-PCR (RT-PCR) was used to identify the BCR-ABL1 transcript. RT-PCR was performed according to European recommendations, as described previously. Results: Clinical assessment confirmed the first chronic phase of a chronic myeloid leukemia. Chromosomal formula showed an homogeneous structural translocation: t(1;9;12;22)(p35-36;q34;p13;q11) involving four chromosomal regions, 1p35-36, 9q34, 12p13 and 22q11.2. Molecular investigation confirmed the Ph nature of the chromosomal abnormality by showing the BCR-ABL1 transcript. The patient was treated by using Imatinib molecule with a favorable therapeutic response. Summary/Conclusion: A new variant Ph chromosome-positive CML in the chronic phase is reported. It is a four-way Ph translocation t(1;9;12;22)(p35-36;q34;p13;q11) involving four chromosomal regions, 1p35-36, 9q34, 12p13 and 22q11.2. Imatinib, the first-generation TKI treatment was efficient.

PB1912: MYELODYSPLASTIC SYNDROME WITH ISOLATED 5Q DELETION AND ASSOCIATED 5Q DELETION TO 13Q DELETION

Background: The 5q deletion is the classic chromosomal abnormality in myelodysplastic syndrome (MDS). Critical regions are 5q31 and 5q33, leading to gene haplodeficiency and other pathogeneic mechanisms. 13q deletion occurs in about 2% of MDS cases. This deletion involved band 13q14. Aims: Here we describe cytogenetic abnormalities in a Tunisian cohort of MDS patients and we shed the light on cases with 5q deletion. Methods: Cytogenetic analyses of bone marrow cells were performed according to standard procedures for 12 Tunisian patients presenting with MDS. Cytogenetic analysis was performed on lymphocytes and bone marrow cells. Cultured cells were treated with colchicine and hypotonic KCl solution. The pellet was fixed and washed in methanol-acetic acid (3:1). The cells were resuspended in fixative and dropped onto slides. Chromosome-banding analysis was performed using RHG and GTG banding techniques. Three karyotypes and 25 metaphases were analyzed according to the International System for Human Cytogenetic Nomenclature. Results: Cytogenetic abnormalities were detected in 9 cases and del5q was recorded in four patients (3 patients with refractory anemia (RA) and a fourth with RA and excess blasts-T (RAEB-T)). The del5q was isolated in two RA cases as a del (5) (q13 q33) in a new diagnosed and a treated patients. It was associated in two patients: a RA patient explored during the evolution, harboring a del (5) (q12 q33) associated to a del (13) (q12 q14) and the RAEB-T patient harbouring a del (5) (q21 q31) associated to a clonal hyperploidy. Summary/Conclusion: The diagnosis of MDS was based on cytomorphology and cytogenetics. However, nowadays, an increasing number of molecular aberrations have been discovered and included in the WHO revised new classification of myeloid neoplasms. The del13q described in MDS seems to be a marker of the evolution of the clonal cells harboring 5q deletion. However, the molecular basis and the implicated genes of the deletion of the long arm of chromosome 13 associated to the 5q deletion clone remain until now not well explored.

Beta Thalassemia and Klinefelter syndrome: a rare occurrence

Backgroundβ-Thalassemia is an inherited haematological blood disorder in the HBB gene, and variations in this HBB gene lead to the absence/deficiency of the Beta chain synthesis of haemoglobin leading to severe anaemia. Klinefelter syndrome is a chromosomal abnormality that affects physical and cognitive development in males. Affected individuals are taller, show gynaecomastia and behavioural problems and have small testes that do not produce much testosterone. We describe a boy with β-Thalassemia major referred for chromosomal analysis due to delayed puberty and short stature. This is a second case reported in the literature that gives information on two different contradicting genetic disorders in a single individual but novel case as he exhibits additional short-stature phenotype.Case presentationA 17-year-old boy with short stature and gonadal dysfunction was referred for chromosomal analysis. He needed blood transfusion every 4 weeks. The GTG banding for chromosomal analysis and standard PCR for variant detection of HBB gene, Bi-directional Sanger sequencing of the PCR products and multiplex PCR for Y microdeletion of the AZF a, b and c regions on the Y chromosome were performed. The cytogenetic analysis revealed a karyotype of 47,XXY. The HBB gene detected two heterozygous variants forming a pathogenic compound heterozygous condition. The multiplex PCR revealed that the AZF a, b and c regions were intact and were not deleted.ConclusionTo our knowledge, this is the second case of a patient with β-Thalassemia associated with Klinefelter syndrome but a novel case with short-stature phenotype association instead of tall stature. The possible association of these two disorders and the unusual phenotypic presentation are discussed. This study highlights the possibility of ruling out Thalassemia while evaluating patients with short stature and delayed puberty.

Cytogenomic Characterization of a Novel de novo Balanced Reciprocal Translocation t(1;12) by Genome Sequencing Leading to Fusion Gene Formation of EYA3/EFCAB4b

Introduction: The accurate detection of breakpoint regions of disease-associated chromosomal rearrangements helps understand the molecular mechanisms and identify the risks involved with disrupted genes. Methods: In this study, a girl with growth retardation is characterized using positional cloning and genome sequencing. The techniques include fluorescence in situ hybridization (FISH) with paint (WCP) and bacterial-artificial chromosomes (BAC) probes, PCR, real-time PCR, and short and long-read sequencing. Results: The translocation was identified by GTG banding and confirmed by WCP FISH. Microarray ruled out the involvement of other copy number variations except for 6 homozygous regions which are not disease-causing variants. Fine mapping with FISH showed split signals with BAC clone RP11-312A3. Genome sequencing of short-read with an average 30× depth and long-read sequencing technology with a 3.8× coverage identified both breakpoints, confirmed by Sanger sequencing, that showed microhomology. The breakpoint at 1p and 12p regions disrupted EYA3 and EFCAB4B genes. Expression analysis of EYA3 showed a 7-fold increase, suggesting the formation of a fusion gene with EFCAB4B. EYA3 is involved in skeleton development, and EFCAB4B plays a role in calcium metabolism, which may be relevant for the patient’s phenotype. Conclusion: The systematic application of genome techniques to translocations and their advantages is discussed.

Correlation of Maternal Age and Chromosomal Abnormality in Products of Conception- A Single Centric Study

Purpose: The purpose of this study is to find out the types and incidence rate of chromosomal abnormalities and the relationship between maternal age and chromosomal abnormality in products of conception by retrospective analysis. Method: Karyotype study using standard GTG banding and FISH study for aneuploidy detection was done from products of conception samples. Results: A total of 513 cases of products of conception were studied retrospectively. 98 cases were studied by conventional cytogenetic technique and 415 cases were studied by a FISH method. The chromosomal abnormality was observed in 97 cases (18.91%). Trisomy was found to be a major chromosomal abnormality amongst the cases studied, followed by triploidy and monosomy. A higher incidence of chromosomal abnormality was found in women with advanced maternal age as compared to the maternal age less than 35 years.